Expression of XCR1 in EOC cell lines. Expression of XCR1 in EOC cell lines. A, cell surface expression of XCR1 in the EOC cell lines using goat anti-human PE-conjugated XCR1 antibodies. Solid thick line – XCR1 expression, solid thin line – negative control (secondary antibody only). Images shown are representative of at least 3 independent experiments. B, expression of XCR1 was tested by immunofluorescent staining of immortalized NOSE and serous EOC cell lines, as indicated. Images were generated using a Zeiss fluorescent microscope and × 40 objective lens. The fluorescent signal from XCR1 staining (red) was overlaid with 4′, 6-diamidino-2-phenylindole (DAPI) staining (blue). A magnified image of XCR1 staining in SKOV-3 converted to a black-and-white image is shown in the lower panel to show membranous immunoreactivity with XCR1 antibodies (white arrows). Intensity of XCR1 staining in cellular protrusions along the dotted line was quantified using the ImageJ (NIH) line scan feature. Bars = 20 μm and 5 μm on the top and bottom, respectively. C, expression of XCR1 was probed in total cell lysates prepared from nonimmortalized NOSE (HOSEpiC), immortalized OSE (T1074), and serous EOC cell lines (IGROV-1 and Caov-3) using goat anti-human XCR1 antibodies. β-Tubulin was used as a loading control. Images were generated using enhanced chemoluminescence application of Chemidoc (Bio-Rad). D, immunohistochemical analysis of XCR1 expression in primary and metastatic specimens of EOC. Representative images of negative and positive cases of primary EOC as well as negative and positive cases of EOC metastasis to the omentum, as indicated. Brown – XCR1, blue – hemotoxylin. Images were generated using the Aperio ScanScope digital slide scanner. Magnification: × 10. Mijung Kim et al. Mol Cancer Res 2012;10:1419-1429 ©2012 by American Association for Cancer Research